Compulsive use of alcohol has been hypothesized to be driven by multiple sources of reinforcement that change with the individual's transition from social drinking to abuse and substance dependence on alcohol. The overarching hypothesis behind the present project is that understanding the dysregulations of the gene regulatory network that underlie the neuroadaptive changes associated with alcohol dependence will allow the identification of new and more effective therapeutic targets for alcohol abuse. Specifically, the goal of the present study is to use a state of the art systems biology strategy in conjunction with a validated rat model of dependence-induced escalated drinking and laser microdissection to reconstruct a Central Nervous System interactome (CNSi). The CNSi will integrate multiple levels of gene expression regulation including the transcriptional (mRNA), post-transcriptional (miRNA) and post-translational (protein-protein interactions and modulators of transcription factor activity) levels. The CNSi will be used to identify the master regulator genes (MRs) that activate specific gene signatures associated with the transition from moderate to excessive alcohol intake associated with dependence in brain regions of the reward and stress systems. The present multiple principal investigator (MPI) project will involve 2 components: 1) The Scripps Research Institute (TSRI) component will carry out the animal models and laser microdissections of brain regions relevant to the reward and stress systems; 2) the Columbia University (CU) component will execute the systems biology strategy. It is expected that the identification of MRs of excessive alcohol intake and modulators of their activity will inform new therapeutic strategies and candidate druggable targets.